JPS5931786A - Phosphatidyl choline-type phospholipid compound - Google Patents
Phosphatidyl choline-type phospholipid compoundInfo
- Publication number
- JPS5931786A JPS5931786A JP14096382A JP14096382A JPS5931786A JP S5931786 A JPS5931786 A JP S5931786A JP 14096382 A JP14096382 A JP 14096382A JP 14096382 A JP14096382 A JP 14096382A JP S5931786 A JPS5931786 A JP S5931786A
- Authority
- JP
- Japan
- Prior art keywords
- compound
- formula
- chloroform
- product
- added
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
- A61K9/1271—Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers
- A61K9/1273—Polymersomes; Liposomes with polymerisable or polymerised bilayer-forming substances
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Dispersion Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicinal Preparation (AREA)
Abstract
Description
【発明の詳細な説明】
この発明はリン脂質化合物に係り、特に、重合性の基を
有するホスファチジルコリン型リン脂質化合物に関する
。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to phospholipid compounds, and particularly to phosphatidylcholine type phospholipid compounds having a polymerizable group.
現在、医薬物質や酵素などを微小なカプセルに封入して
医薬品として提供する試みが種々なされている。Currently, various attempts are being made to encapsulate medicinal substances, enzymes, etc. into minute capsules and provide them as pharmaceuticals.
このようなマイクロカブセル化の初期の方法は、乳化法
による高分子化合物のカプセル化(特公昭45−275
7号および特公昭50−17950号)や界面重縮合反
応による重合体(ポリアミドなど)の生成を伴なったカ
プセル化(特公昭53−7395号および特公昭53−
7396号)である。しかしながら、これらの方法では
、例えばカプセル化材料である重合体の著しb毒性やそ
の合成過程で必要な有機溶媒(例えば、ジオキサン、ア
セトン、キシレン、ベンゼン)がカプセル中へ残存する
ことによる毒性、さらKはカプセルの粒径が大きい(数
μm〜1000μm)ために血栓などの障害を引き起し
易因という問題がちり、得られたカプセルを医薬品とし
て使用するには問題があった。The early method of microencapsulation was the encapsulation of polymer compounds by emulsification method (Japanese Patent Publication No. 45-275).
7 and Japanese Patent Publication No. 50-17950) and encapsulation accompanied by the production of polymers (polyamide, etc.) by interfacial polycondensation reaction (Japanese Patent Publication No. 7395 and Japanese Patent Publication No. 1983-1983)
No. 7396). However, in these methods, for example, the toxicity of the polymer that is the encapsulating material, the toxicity due to the organic solvents (e.g. dioxane, acetone, xylene, benzene) required in the synthesis process remaining in the capsule, Sarak has a large particle size (several micrometers to 1000 micrometers) of its capsules, which tends to cause problems such as thrombosis, making it difficult to use the resulting capsules as pharmaceuticals.
ところで、医薬物質や酵素をマイクロカプセルに封入す
る主たる目的は、主として、生体内で不安定な医薬物質
や酵素の活性を長時間保持させ、その効果を長時間持続
させることである。By the way, the main purpose of encapsulating pharmaceutical substances and enzymes in microcapsules is to maintain the activity of pharmaceutical substances and enzymes that are unstable in vivo for a long period of time, and to maintain their effects for a long period of time.
したがって、特に医薬品としてマイクロカプセルを使用
することを目指す場合、既述の従来技術の問題点を含め
て次の条件が特に重要となる。すなわち、
(1) マイクロカプセル粒径をより微小化(数μm
以下)すること、
(2)生体内におけるカプセルの安定性を向上させるこ
と、および
(3) カプセル材料の毒性を低下させることである
。Therefore, especially when aiming to use microcapsules as a pharmaceutical, the following conditions are particularly important, including the problems of the prior art described above. (1) Making the microcapsule particle size even smaller (several μm)
(2) improve the stability of the capsule in vivo; and (3) reduce the toxicity of the capsule material.
このような条件をかなりの程度に満足する技梠
術として、生体膜の成分である各種のリン葉質が水中で
微小な球状の集合体(リポソーム、粒径0.05〜10
μm)を形成することを利用して、重合性の基を有する
リン脂質化合物を合成し、これによって形成されるリポ
ソームをそのままの状態を保って重合し、マイクロカプ
セルを得る技術が、最近、報告されている。例えば、S
、IL、 Regen他によるJournal of
the AmericanChemical 5oci
ety 、 104 、791〜795(1982)(
以下、文献(1)という) 、H−H−Hub他による
Angewanta Chemle Internat
ionalEdit1on英語版、19,938〜94
0(1980)(以下、文献(II)という)、および
A、AkimotoらによるAngewante Ch
emleInternational Edition
英語版、20.90〜91 (1981)(以下、文献
価という)参照。As a technology that satisfies these conditions to a considerable extent, various phosphorus substances, which are components of biological membranes, are formed into minute spherical aggregates (liposomes, particle size 0.05 to 10 mm) in water.
Recently, a technology to obtain microcapsules by synthesizing a phospholipid compound having a polymerizable group and polymerizing the resulting liposomes while maintaining the same state has been reported. has been done. For example, S
, IL, Journal of Regen et al.
the American Chemical
ety, 104, 791-795 (1982) (
(hereinafter referred to as document (1)), Angewanta Chemle International by H-H-Hub et al.
ionalEdit1on English version, 19,938-94
0 (1980) (hereinafter referred to as Document (II)), and Angewante Ch by A. Akimoto et al.
emleInternational Edition
See English version, 20.90-91 (1981) (hereinafter referred to as literature value).
これら文献(I)〜(2)には次に示す重合性リン脂質
化合物が記載されている。These documents (I) to (2) describe the following polymerizable phospholipid compounds.
文献(I)
および
文献(II)
一5=
および
1
−
文献(1)
1
〇−
これら合成リン脂質化合物は水中でリポソーム(あるい
はベヒクル)を形成する。このリポソームをそのtまの
状態でマイクロカプセル化させるには、各化合物におけ
る重合性の基(す6一
なわち、文献(1)の化合物ではビニル基、文献(ID
の化合物ではジアセチレン基、文献価の化合物ではブタ
ジェン基)を重合させる必要がある。Reference (I) and Reference (II) 15= and 1 - Reference (1) 1 0- These synthetic phospholipid compounds form liposomes (or vehicles) in water. In order to microencapsulate this liposome in its original state, the polymerizable groups in each compound (i.e., the vinyl group in the compound of literature (1), the vinyl group in the compound of literature (1),
It is necessary to polymerize the diacetylene group in the compound of 1, and the butadiene group in the compound with literature value.
文献(Dの化合物の重合を開始させるには、適当な重合
開始剤(例えば、アゾビスインブチロニトリル等のアゾ
系開始剤、過酸化ベンゾイル等の有機過酸化物系開始剤
、過硫酸カリウム等の開始剤)を加えることが必要であ
る。したかって、文献(1)の化合物は、これを重合さ
せた場合、重合開始剤断片が残存し、その毒性が問題と
なってくる。なお、この化合物は紫外線照射によっても
重合可能であるが高いエネルギーを必要とし、よシ短波
長光を必要とする。Literature (To initiate the polymerization of the compound of Therefore, when the compound of document (1) is polymerized, polymerization initiator fragments remain, which poses a problem of toxicity. Although this compound can be polymerized by ultraviolet irradiation, it requires high energy and requires very short wavelength light.
文献(n)および[相]の化合物は紫外線の照射により
重合するので、文献(1)の化合物にみられるような問
題は生じない。しかしながら、その重合丈メh大’ (
[) σ)イビ、パ≧々勿 7“111、−CH=CH
−CH=CH−重合 −C)(−CH=CH−CH−/
等の形式で進行し、得られるカブセル比重合体中に多数
の不飽和結合が残る。不飽和結合部位は生体内において
代謝を受は易く、その代謝物(ビニル基のエポキシ化物
等)が著しく毒性の高いものとなる可能性がある。Since the compounds in Document (n) and [Phase] are polymerized by irradiation with ultraviolet rays, the problems seen in the compound in Document (1) do not occur. However, the polymerization length is large (
[) σ) Ibi, pa ≧ and of course 7"111, -CH=CH
-CH=CH- polymerization proceeds in the form -C) (-CH=CH-CH-/, etc., and many unsaturated bonds remain in the resulting capsule specific polymer. The unsaturated bond sites are difficult to metabolize in vivo. The metabolites (epoxidized vinyl groups, etc.) may be highly toxic.
したがって、この発明の目的は、重合開始剤の添加を必
要とせずに重合してマイクロカプセル化することができ
、しかも重合して得られるマイクロカプセル重合体中に
不飽和結合が残存しないタイプの合成リン脂質化合物を
提供することである。Therefore, an object of the present invention is to synthesize a type of polymer that can be polymerized and microencapsulated without the need for the addition of a polymerization initiator, and in which no unsaturated bonds remain in the microcapsule polymer obtained by polymerization. An object of the present invention is to provide phospholipid compounds.
この発明罠よれば、一般式
%式%
の正の整数、およびtは13ないし21の正の整数)で
示されるホスファチジルコリン型リン脂質化合物が提供
される。According to this invention, there is provided a phosphatidylcholine type phospholipid compound represented by the general formula %, where t is a positive integer from 13 to 21.
一般式囚で示されるこの発明のリン脂質化合物を製造す
るには、まず一般式
(ただし、mおよびnは既述のとおり)で示される酸ク
ロリドを一般式
(ただし、Lは既述のとおシ)で示されるグリセロール
誘導体と縮合反応させて一般式を得る。この縮合は式(
B)と式(9)の化合物モル比0.5:1ないし3:1
の割合で用い、適当量の脱塩酸剤(ピリジン等)を含有
する適当な溶媒(例、tば、無水のテトラヒドロフラン
、クロロホルム、ジクロルメタン等)中で0℃ないし3
0℃の温度でおこなう。反応時間は、通常、工ないし2
0時間である。To produce the phospholipid compound of the present invention represented by the general formula, first, an acid chloride represented by the general formula (where m and n are as described above) is converted into an acid chloride represented by the general formula (where L is as described above). The general formula is obtained by condensation reaction with the glycerol derivative shown in c). This condensation is expressed by the formula (
B) and the compound of formula (9) molar ratio 0.5:1 to 3:1
in an appropriate solvent (e.g., tungsten, anhydrous tetrahydrofuran, chloroform, dichloromethane, etc.) containing an appropriate amount of a dehydrochlorination agent (pyridine, etc.) at 0°C to 3°C.
Perform at a temperature of 0°C. The reaction time is usually between 1 and 2 hours.
It is 0 hours.
ついで、式(ハ)で示される縮合生成物をVan R。Then, the condensation product represented by formula (c) was subjected to VanR.
H1rt他によってPharm、 Acts H@lv
、 + 33 。Pharm, Acts H@lv by H1rt et al.
, +33.
349 (1958)に記載されている方法に準じて次
に示す工程で反応させることによって目的とする一般式
(4)の化合物が得られ石。The target compound of general formula (4) can be obtained by reacting in the following steps according to the method described in 349 (1958).
10−
縮合生成物(D)+Ct2−POCH’2CH2Br(
2)A安CH3COO−
なお、式(B)で示される酸クロリドのうち、mが0の
ものは、Y、 Iwakura他によってBull。10- Condensation product (D) + Ct2-POCH'2CH2Br(
2) Amino CH3COO- Among the acid chlorides represented by the formula (B), those in which m is 0 were described as Bull by Y. Iwakura et al.
Chem、 Soc、 、 Jap、 + 4±、18
6〜191(1968)に記載されている方法に従って
p−カルボキシスチレンと塩化チオニルとを反応させる
ことによって合成できる。また、式(B)で示される酸
クロリドのうちmが1の場合は、まずX3.8van他
によってOrganlc 5yntheaLs+2.2
76(1950)に記載された方法を改良して得られる
式
%式%
()
(ただし、nは既述のとおり、Rはメチル基、エチル基
、プロピル基捷たはブチル基)で示されるアルカンジカ
ルボン酸モノエステルeモノクロリドを無水塩化アルミ
ニウムの存在下に臭化エチルベンゼンと反応させて式
(ただし、nおよびRは既述のとおシ)で示される化合
物を得る。ついでこれを過剰のアルカリ(例えば、水酸
化ナトリウム、水酸化カリウム)とアルコール中で加熱
反応させた後、塩酸等で酸性化して式
で示される化合物を得る。これを塩化チオニルまたはシ
ーウ酸クロリド等と反応させることによって式(B)で
示される化合物が得られる。Chem, Soc, , Jap, + 4±, 18
6-191 (1968) by reacting p-carboxystyrene and thionyl chloride. In addition, when m is 1 in the acid chloride represented by the formula (B), first, Organlc 5yntheaLs+2.2
76 (1950) is obtained by improving the method described in % Formula % () (where n is as described above, R is a methyl group, an ethyl group, a propyl group, or a butyl group). Alkanedicarboxylic acid monoester e monochloride is reacted with ethylbenzene bromide in the presence of anhydrous aluminum chloride to obtain a compound represented by the formula (where n and R are as defined above). Next, this is reacted with excess alkali (eg, sodium hydroxide, potassium hydroxide) under heating in alcohol, and then acidified with hydrochloric acid or the like to obtain a compound represented by the formula. A compound represented by formula (B) can be obtained by reacting this with thionyl chloride, sioyl chloride, or the like.
一般式(C)で示されるグリセロール誘導体はり。A glycerol derivative beam represented by general formula (C).
Arnoldらによって、Lieb、 Ann、 Ch
em、e 703 t234〜239 (1967)に
記載されている方法を改良して、2−フェニル−5−m
−ジオキkyルキルプロミドを反応させた後、エタノー
ル中で硫酸により加水分解することによって得られる。By Arnold et al., Lieb, Ann, Ch.
Em, e 703 t234-239 (1967) was modified to produce 2-phenyl-5-m
- Obtained by reacting dioxykyl bromide and then hydrolyzing it with sulfuric acid in ethanol.
なお、一般式(C)で示されるグリセロール誘導体のう
ちtが13.15,17.19または21のものを合成
するための原料であるアルキルプロミドは市販品として
安価に入手できるので都合がよい。Note that among the glycerol derivatives represented by general formula (C), alkyl bromide, which is a raw material for synthesizing those with t of 13.15, 17.19, or 21, is convenient because it can be obtained at low cost as a commercial product. .
この発明のホスファチジルコリン型リン脂質化合物は、
天然リン脂質からリポソームを得るための一般的な方法
(例えば、D、Papahadjopouloa他によ
るBloahimla et Biophyaia A
eta、 13 L639〜652 (j967)等参
照)に、準じてその1種または2種以上を処理すること
によって水中テ粒径0.03〜1μmのリポソームを形
成する。リポソームの形成のしやすさは、一般式(3)
においてmが0のときよりもmが1のときの方13−
が大きく、また、nの値は大きい方がよい。こうして得
られるす列?ンーム分散水溶液に紫外光全光や紫外単色
光(例えば、240 nmt 310nm)を照射する
ことによって迅速に重合反応が進行し、マイクロカプセ
ルが生成する。この重合に際して重合開始剤等の添加剤
を加える必要は一切ないとともに、既述の文献(If)
および(至)における化合物のように重合体中に不飽和
結合が残存することはない。この発明のホスファチジル
コリン型リン脂質化合物はそれ自体の毒性も低い。The phosphatidylcholine type phospholipid compound of this invention is
General methods for obtaining liposomes from natural phospholipids (e.g. Bloahimla et Biophyaia A by Papahadjopouloa et al.
eta, 13 L639-652 (j967), etc.), liposomes having an in-water particle size of 0.03 to 1 μm are formed by treating one or more of them in accordance with the method described in 13 L639-652 (j967). The ease of forming liposomes is determined by the general formula (3)
13- is larger when m is 1 than when m is 0, and the value of n is preferably larger. The column obtained in this way? By irradiating the aqueous solution of ultraviolet light or monochromatic ultraviolet light (for example, 240 nm to 310 nm), a polymerization reaction rapidly proceeds to produce microcapsules. There is no need to add any additives such as a polymerization initiator during this polymerization, and there is no need to add any additives such as a polymerization initiator.
Unlike the compounds in and (to), no unsaturated bonds remain in the polymer. The phosphatidylcholine type phospholipid compound of this invention also has low toxicity itself.
したがって、この発明のホスファチジルコリン型リン脂
質から得られるマイクロカプセルは゛その中に医薬物質
や酵素等を封入(医薬物質や酵素等を含有する生理的に
許容できる水溶液中での重合による)することによって
医薬品として、あるいは臨床検査等各種検査用試薬とし
て有用である。また、写真用感光剤等や色素等を封入す
ることによってよシ鮮明々写真や印刷画像を得るための
材料ともなる。Therefore, the microcapsules obtained from the phosphatidylcholine type phospholipids of the present invention can be used to produce pharmaceuticals by encapsulating a pharmaceutical substance, enzyme, etc. therein (by polymerization in a physiologically acceptable aqueous solution containing the pharmaceutical substance, enzyme, etc.). It is useful as a reagent for various tests such as clinical tests. In addition, by encapsulating photographic photosensitizers, dyes, etc., it can also be used as a material for obtaining sharper photographs and printed images.
14− 以下、この発明の実施例を記す。14- Examples of this invention will be described below.
実施例1
(A−1) H,Hib’bert他によるJ、
Amer、 Chem。Example 1 (A-1) J by H, Hib'bert et al.
Amer, Chem.
Soc、、62.160:l〜1613(1929)に
記載のによるJun、 L[eb、 Ann、 Che
m、 + 709 +234〜239 (1967)に
記載の方法に従って臭化ステアリルと反応させて、2−
ステアリロキシ−1,,3−fロノクンジオール金合成
した。Soc, 62.160:1-1613 (1929) by Jun, L [eb, Ann, Che
2-
Stearyloxy-1,,3-f lonocundiol gold was synthesized.
C73,75(73,3)
ただし、()内の値はC21T(4403の計算値
5 31、.9
6 26.1
20 22.7
21 14、I
IRスペクトル(KBrベレット)(cm−1):33
50.2940.
2870.1470.
1120.1080、
80
(A−2) 2−ステアリロキシ−1,3−プロノぐ
フジオール4.55グラム(13,2ミリモル)に無水
クロロホルム30−および乾iビlJジン2.5ml
f加えて溶解し、これにY、 Iwakuraらによる
Bull、Chom、Soc、+ Jap−s ↓1
,186〜191(1968)に記載の方法に従って合
成したp−ビニルベンゾイルクロリド2.2グラム(1
3,2ミリモル)の無水クロロホルム(15d)溶iを
30分間かけて徐々に滴下した。この反応溶液を室温で
18時間攪拌した後、これを0.5N塩酸各50−で2
回、水各50−で3回、炭酸ナトリウム1チ水溶液50
Tnlで1回洗浄し、硫酸す) IJウムで脱水した。C73,75 (73,3) However, the values in parentheses are C21T (calculated value of 4403 5 31,.9 6 26.1 20 22.7 21 14, I IR spectrum (KBr pellet) (cm-1) :33
50.2940. 2870.1470. 1120.1080, 80 (A-2) 4.55 grams (13.2 mmol) of 2-stearyloxy-1,3-pronogfudiol in 30-ml of anhydrous chloroform and 2.5 ml of dry bicarbonate.
Add f and dissolve, to which Y, Bull, Chom, Soc, + Jap-s by Iwakura et al. ↓1
, 186-191 (1968).
3.2 mmol) of anhydrous chloroform (15d) was gradually added dropwise over 30 minutes. After stirring the reaction solution at room temperature for 18 hours, it was diluted with 0.5N hydrochloric acid for 2 hours.
3 times with 50% water each, 1% sodium carbonate aqueous solution 50%
Washed once with Tnl and dehydrated with IJ sulfuric acid.
得られた溶液をろ過し、減圧下で溶媒を除去した。残分
をシリカゲルカラム(直径4.5 crn %長さ35
crrL)分画に供し、目的分画を集め、溶媒を減圧留
去した後、油状残分をオイルポンプを用いて50℃で乾
燥して目的物を得た。収量3.6グラム・シリカダル薄
層クロマトグラフィー(TLC)(W媒りロロホルム)
:Rf=0.23(紫外線吸収、工2による呈色あり)
質量スペクトル: M+=474 (分子量474)1
7−
元素分析値: C76,07(75,9)Hlo、60
(10,5)
ただし、()内はC30H5004の計算値。The resulting solution was filtered and the solvent was removed under reduced pressure. Transfer the residue to a silica gel column (diameter 4.5 crn, length 35
crrL) fractionation, the desired fractions were collected, the solvent was distilled off under reduced pressure, and the oily residue was dried at 50° C. using an oil pump to obtain the desired product. Yield: 3.6 grams Silica Dull Thin Layer Chromatography (TLC) (Roloform in W medium)
:Rf=0.23 (ultraviolet absorption, coloring due to engineering 2) Mass spectrum: M+=474 (molecular weight 474) 1
7- Elemental analysis value: C76,07 (75,9) Hlo, 60
(10,5) However, the values in parentheses are calculated values for C30H5004.
”C−NMRスペクトルδ(ppm)値:6
26.1
20 23.7
21 14.1
22 166.2
23 128.9
24 129.9
25 126.1
26 142.1
27 135.9
28 116.5
18−
ロモエチル)エステルノ合成
Von R,HlrtらによるPharma、 Act
a He1v−+lユ、349 (1958)に記載の
方法に従って合成したCl3F(0)CH2CH2Br
5. Oグラムの無水クロロホルム(15rn1.)
溶液を氷冷し、これにトリエチルアミン5−1および上
記(4)で得たモノオール1.5グラムのクロロホルム
(101nl)溶液を2時間かけて別々のロートから同
時に滴下した。この反応溶液を室温で2時間反応させた
後、TLCで反応の進行を確認した。ついで、この反応
溶液を氷冷し、これに0.5N塩化カリウム水溶液20
−を1時間かけて滴下した。さらに1時間攪拌した後メ
タノール15+dを加え、クロロホルム層(暗赤色)を
分液した。クロロカダルカラムクロマトグラフィー(ク
ロロホルム/メタノール= 10/1 )で目的物(黄
色油状)を単離した。収量1,4グラム。"C-NMR spectrum δ (ppm) value: 6
26.1 20 23.7 21 14.1 22 166.2 23 128.9 24 129.9 25 126.1 26 142.1 27 135.9 28 116.5 18- Lomoethyl) ester synthesis Von R, Hlrt et al. Pharma, Act
a Cl3F(0)CH2CH2Br synthesized according to the method described in He1v-+lU, 349 (1958)
5. O grams of anhydrous chloroform (15rn1.)
The solution was cooled with ice, and a solution of triethylamine 5-1 and 1.5 g of the monool obtained in (4) above in chloroform (101 nl) was simultaneously added dropwise from separate funnels over 2 hours. After reacting this reaction solution at room temperature for 2 hours, the progress of the reaction was confirmed by TLC. Next, this reaction solution was cooled on ice, and 200% of a 0.5N aqueous potassium chloride solution was added to it.
- was added dropwise over 1 hour. After further stirring for 1 hour, 15+d methanol was added and the chloroform layer (dark red) was separated. The target product (yellow oil) was isolated by chlorocadal column chromatography (chloroform/methanol = 10/1). Yield 1.4 grams.
19−
り呈色あり)
15C−を収Rスペクトルδ値ニ
ア〜19 29.7
20 22.7
21 14.1
22 165.8
23 128.8
24 130.0
25 126.1
26 142.1
27 135.9
28 116.6
2O−
IRスペクトル(KBrベレットXcrn−1):29
30.2850.1720.1610,1460゜12
70.1210.1100,1015.99o1910
.860
の合成
前記(B)で得たリン酸エステル化合物1.3グラムを
無水ブタノン15−に溶解し、ドライアイス−メタノー
ル浴でドラッグして冷却し液化したトリメチルアミン1
5fnlを加えてステンレス管中に入れて封管し、55
℃に熱し、攪拌しながら10時間反応させた。管を開放
し、これを0℃に冷却し、生じた沈でん(生成物)?:
ろ集し、アセトンで洗浄した後乾燥した。この生成物か
ら臭素イオンを除去するためこれをメタノール50−一
水5−の混合溶媒に懸濁させAg(0OCCH5)2ダ
ラムを加え、暗所で攪拌しながら2時間反応させた。反
応溶液をろ過し、ろ液を濃縮した後、シリカダルカラム
クロマトグラフ=21−
イー(クロロホルム/メタノール/水=65/2515
)分画に供し、目的分画を集め、濃縮した。この濃縮
物をクロロホルムに溶解し、多量のアセトン中に注下、
再沈でんさせ、沈でんを遠心分離し、真空乾燥して目的
とするホスファチジルコリン型リン脂質化合物を得た。19- with red coloring) 15C- was collected R spectrum δ value near ~ 19 29.7 20 22.7 21 14.1 22 165.8 23 128.8 24 130.0 25 126.1 26 142.1 27 135.9 28 116.6 2O- IR spectrum (KBr pellet Xcrn-1): 29
30.2850.1720.1610,1460°12
70.1210.1100, 1015.99o1910
.. Synthesis of 860 1.3 grams of the phosphoric acid ester compound obtained in the above (B) was dissolved in anhydrous butanone 15-, and the solution was cooled and liquefied by dragging it in a dry ice-methanol bath.
Add 5fnl and put it in a stainless steel tube and seal it.
The mixture was heated to 0.degree. C. and reacted for 10 hours with stirring. Open the tube, cool it to 0°C, and collect the resulting precipitate (product). :
It was collected by filtration, washed with acetone, and then dried. In order to remove bromide ions from this product, it was suspended in a mixed solvent of 50 parts of methanol and 5 parts of water, Ag(0OCCH5)2 Durum was added, and the mixture was reacted for 2 hours with stirring in a dark place. After filtering the reaction solution and concentrating the filtrate, silica column chromatography = 21-E (chloroform/methanol/water = 65/2515
) The desired fractions were collected and concentrated. This concentrate was dissolved in chloroform and poured into a large amount of acetone.
The precipitate was reprecipitated, centrifuged, and vacuum dried to obtain the desired phosphatidylcholine type phospholipid compound.
22−
”C−NMRス被クトり(δ値)ニ
ア〜19 29.7
20 22.7
21 14.1
22 166.1
23 129.1
24 129.9
25 126.0
26 142.0
27 135.9
28 116.5
29 59.4
30 66.3
31 54.3
1715.1615.1470.128へ1210.1
090.1060.970.920.860,820
参考例
実施例1で得たリン脂質化合物を濃度が3.5X10M
となるように水に加えた。これを窒素下で超音波処理(
75W、40℃で5分間)シ、さらに室温で1時間放置
した後、500Wの水銀灯(牛尾電機製UI−501C
型)を用い、光路長30cIrLで光照射した。リン脂
質化合物の紫外吸収スペクトルを経時的に測定した。22- "C-NMR spectrum coverage (δ value) near ~ 19 29.7 20 22.7 21 14.1 22 166.1 23 129.1 24 129.9 25 126.0 26 142.0 27 135 .9 28 116.5 29 59.4 30 66.3 31 54.3 1210.1 to 1715.1615.1470.128
090.1060.970.920.860,820 Reference Example The phospholipid compound obtained in Example 1 was added at a concentration of 3.5×10M.
Added to water so that This was treated with ultrasonication under nitrogen (
75W at 40℃ for 5 minutes), and then left at room temperature for 1 hour, then heated with a 500W mercury lamp (UI-501C manufactured by Ushio Electric).
Light was irradiated with an optical path length of 30 cIrL. The ultraviolet absorption spectra of phospholipid compounds were measured over time.
反応に伴い、特性吸収帯(λm□260 nm付近)の
吸収が減少し、約15分後に該吸収が消失し、重合が完
了したことを確認した。この重合物の形状を電子顕微鏡
(日立製H−500型)によって測定したところ、平均
粒径0.25μmの微小球形であることがわかった。こ
の重合物液に等溶積のエタノールを加えたところ、その
形状は変化せず、重合マイクロカプセルが重合前に較べ
て安定であることがわかった。また、重合前のリポソー
ム(ベヒクル)の脂質二重層の熱転移温度は約−4,5
℃であったが、重合後では約16.5℃であった。As the reaction progressed, absorption in a characteristic absorption band (around λm□260 nm) decreased, and the absorption disappeared after about 15 minutes, confirming that the polymerization was complete. When the shape of this polymer was measured using an electron microscope (Hitachi Model H-500), it was found to be microspherical with an average particle size of 0.25 μm. When an equal volume of ethanol was added to this polymer solution, its shape did not change, indicating that the polymerized microcapsules were more stable than before polymerization. Furthermore, the thermal transition temperature of the lipid bilayer of liposomes (vehicles) before polymerization is approximately -4.5
The temperature was approximately 16.5°C after polymerization.
実施例2
S、Swam他によるOrganle 5ynthes
is 、 2 +276 (1950)に記載されてい
る方法に従って合成したセパシン酸モノエチルエステル
・モノクロリド10.0グラム(40,2ミリモル)、
ブロモエチルベンゼン6.2グラム(33,5ミIJミ
ニウム11.2グラム(83,8ミリモル) ’c 添
加した。この混合物を水浴中で4時間攪拌した後、室温
で終夜攪拌した。水浴中で再度冷却した後、この混合物
に氷と水を添加し、ついで濃塩酸5.7d’iゆっくり
と加えた。これに、メチレンクロリドと水を適当量加え
て振盪した後メ25−
チレンクロリド層を分離し、また水層にメチレンクロリ
Pを加えてさらに抽出した。メチレンクロリド層を合せ
ナトリウムで脱水した後、メチレンクロリドおよびニト
ロベンゼンを減圧下で蒸発除去し、シリカゲルカラム(
i径4.、長さ35儂)で分画し、所望の生成物を得た
。Example 2 Organle 5ynthes by S. Swam et al.
10.0 g (40.2 mmol) of cepacic acid monoethyl ester monochloride synthesized according to the method described in J.I.S., 2 +276 (1950),
6.2 g of bromoethylbenzene (11.2 g (83,8 mmol) of 33,5 mmol) was added. The mixture was stirred in a water bath for 4 hours and then stirred overnight at room temperature. After cooling, ice and water were added to the mixture, and then 5.7 d'i of concentrated hydrochloric acid was added slowly.To this, appropriate amounts of methylene chloride and water were added, and after shaking, the me25-ethylene chloride layer was separated. Then, methylene chloride P was added to the aqueous layer for further extraction.The methylene chloride layers were combined and dehydrated with sodium, and then methylene chloride and nitrobenzene were removed by evaporation under reduced pressure.
i diameter 4. , length 35 degrees) to obtain the desired product.
収量6.6グラム(収率49チ)。Yield: 6.6 grams (yield: 49 g).
質量ス(クトル: M+=396(C2oH2903B
r1の分子量396.398)
チモールブルーによ
る呈色あシ)
1370.1260.1220.1180.1130、
1095.1030
元素分析値:C60,20(60,5)H7,61(7
,4)
ただし、()内のイ直はC2oH2903Br1の計算
値
26−
S5cmN皿ス被クトルδ値:
1 33.1
2 39.0
3 143.9
4 128.4
5 128.8
6 135.8
7 199.8
8 38.5
9.14 24.9,24.3
10〜13 29.3,29.1
15 34.3
16 173.7
17 60.1
18 14.3
27−
(B)p−(クロロカルがニルノナノイル)スチレンの
合成
CB−1) j二記(4)で得たp−(エトキシカル
ビニルノナノイル)フェニルエチルプロミド3.3グラ
ム(83ミリモル)を1級エタノール40m1と粉末水
酸化カリウム6グラムとの混合物に加え、これを還流下
に2.5時間熱した後水冷した。Mass S(ctor: M+=396(C2oH2903B
Molecular weight of r1 396.398) Coloration by thymol blue) 1370.1260.1220.1180.1130,
1095.1030 Elemental analysis value: C60,20 (60,5) H7,61 (7
, 4) However, the values in parentheses are the calculated value of C2oH2903Br1 26- S5cmN dish space vector δ value: 1 33.1 2 39.0 3 143.9 4 128.4 5 128.8 6 135.8 7 199.8 8 38.5 9.14 24.9, 24.3 10-13 29.3, 29.1 15 34.3 16 173.7 17 60.1 18 14.3 27- (B)p -Synthesis of (chlorocal is nilnonanoyl)styrene CB-1) 3.3 g (83 mmol) of p-(ethoxycarvinylnonanoyl) phenylethyl bromide obtained in J.2 (4) is mixed with 40 ml of primary ethanol and powder. Added to a mixture with 6 grams of potassium hydroxide, this was heated under reflux for 2.5 hours and then cooled with water.
生成した沈でんをろ集し、乾燥した後、水10〇−に溶
解した。この溶液に濃塩酸5ゴを加え、生成した沈でん
を集め、減圧乾燥した。この生成物を熱トルエン50−
と石油エーテル150−の混合物から再結晶させて所望
の生成物(p−(ヒドロキシカルボニルノナノイル)ス
チレン)を得た。収量1.1グラム(収率46チ)。The produced precipitate was collected by filtration, dried, and then dissolved in 100 ml of water. Concentrated hydrochloric acid was added to this solution, and the resulting precipitate was collected and dried under reduced pressure. This product was mixed with hot toluene at 50-
and petroleum ether 150 to give the desired product (p-(hydroxycarbonylnonanoyl)styrene). Yield: 1.1 grams (yield: 46 inches).
13C−NMRスペクトル(δ値)ニ
ー9只−
1116,59,1424,4,24,63141,8
4126,31534,1
5128,416180,1
6136,1
7200,0
838,5
1,34(S’、 8H、−(CH2→T)2.31(
d、4H,−C(0)CCI(2−)2.35(t、2
H,−CI42C(0)−0−)5.49(q、2H,
−C=CH2)
29−
2860.1725.1685.1610.1470.
1440.1410.1380.1310.1245.
1190,1120゜995.980.910.840
質量ス被りトル: M+=288 (C1aHz405
の分子量288)
(B−2) 上記(B−1)で得たp−(ヒドロキシ
カルがニルノナノイル)スチレ:/ 1.0グラム(3
,5ミリモル)に乾燥ベンゼン25−およびジメチルホ
ルムアミド1滴を加えて攪拌した。(S') , 8H, -(CH2→T)2.31(
d,4H,-C(0)CCI(2-)2.35(t,2
H, -CI42C(0)-0-)5.49(q,2H,
-C=CH2) 29- 2860.1725.1685.1610.1470.
1440.1410.1380.1310.1245.
1190,1120゜995.980.910.840 Mass overburden torque: M+=288 (C1aHz405
Molecular weight: 288) (B-2) p-(hydroxycal is ninonanoyl) styrene obtained in (B-1): / 1.0 g (3
, 5 mmol) were added with 25 cm of dry benzene and 1 drop of dimethylformamide and stirred.
これにシュウ酸ジクロリド1m(11,8ミリモル)を
滴下し、室温で2時間攪拌した後減圧下で溶媒を除去し
た。残分をオイルlンプで乾燥し、所望の酸クロリド生
成物であるp−(クロロカルボニルノナノイル)スチレ
ンを得た。To this was added dropwise 1 m (11.8 mmol) of oxalic acid dichloride, and after stirring at room temperature for 2 hours, the solvent was removed under reduced pressure. The residue was dried in an oil pump to obtain the desired acid chloride product, p-(chlorocarbonylnonanoyl)styrene.
30−
実施例1 (A−1)で得た2−ステアリロキシ−1,
3−プロノ臂ンジオール0995グラム(2,89ミリ
モル)に乾燥メチレンクロリド30−および乾燥ピリジ
ン0.5−を加えて攪拌した。これに、前記(B−2)
で得た酸クロリド全量(3,5ミリモル)全乾燥メチレ
ンクロリド10−に溶解した溶液を10分間かけてゆっ
くりと滴下し、終夜攪拌した。この反応混合物からメチ
レンクロリド全留去し、残分をクロロホルムに溶解した
。この溶液を0.5N塩酸、水、2チ炭酸ナトリウム水
溶液および水で順次洗浄した後硫酸ナトリウムで脱水し
た。これを減圧下で蒸発処理に供し、残分を乾燥した後
、ベンゼン/エーテル−9/1の流出溶媒を用いてシリ
カダルカラム(直径2CrrL1長さ30儒)で精製し
て所望のエーテル生成物を得た。30- Example 1 2-stearyloxy-1 obtained in (A-1),
To 0,995 grams (2,89 mmol) of 3-prononodiol were added 30 grams of dry methylene chloride and 0.5 grams of dry pyridine and stirred. In addition, the above (B-2)
A solution of the acid chloride obtained above (3.5 mmol) dissolved in 10-10% dry methylene chloride was slowly added dropwise over 10 minutes, and the mixture was stirred overnight. All methylene chloride was distilled off from this reaction mixture, and the residue was dissolved in chloroform. This solution was washed successively with 0.5N hydrochloric acid, water, an aqueous solution of di-sodium carbonate, and water, and then dehydrated with sodium sulfate. This was subjected to evaporation treatment under reduced pressure, and the residue was dried and purified with a silica dull column (diameter 2CrrL, length 30F) using a benzene/ether-9/1 effluent solvent to obtain the desired ether product. I got it.
3l−
Rf=0.28(シクロヘキサ乙4詐酸エチル−3/1
)
(紫外m吸収、プロムチトルカ−に
よる呈色あり)
質量スペクトル:M+−614(分子量614)元素分
析値: C76,38(76,2)810.82(10
,7)
ただし、()の値はC39H6605の計算値
”C−NMRス4クトル(δ値)ニ
ーqワ−
362,014’ 24.91’ 1
16.4 15’ 34.22’
135.9 16’ 173.63
’ 141.8 1“ 14.14
’ 126.2 2“ 22.75
’ 128.4 3“ 31.96
’ 136.1 17“ 26.17
’ 199.7 18“ 7o、58
’ 38.5
9’ 24.3
1690.1610.1475.1400゜1370.
1295.123011185.1135.995.9
80.91o1
840.730
33−
ホスホリルコリンの合成
実施例1 (B)で用いたC42P(0)OCH2CH
2Br 3. Oグラムを乾燥クロロホルム10−に溶
解し、氷冷した。これに、上記((1’lで得たエーテ
ル生成物1.0グラム(29ミリモル)を乾燥クロロホ
ルム10ゴに溶解した溶液、および乾燥トリエチルアミ
ン4−を乾燥クロロホルム1.0tntK:溶解した溶
液をそれぞれ別々のろう斗から同時に滴下した。この反
応混合物を室温で20時間攪拌した後、Q、5M塩化カ
リウム水溶液10−をゆっ〈シ滴下し、さらに室温で1
時間攪拌した。3l- Rf = 0.28 (Cyclohexa Otsu 4 ethyl sulfate - 3/1
) (Ultraviolet m absorption, coloration by promthitorcar) Mass spectrum: M+-614 (molecular weight 614) Elemental analysis value: C76,38 (76,2) 810.82 (10
,7) However, the value in parentheses is the calculated value of C39H6605.
16.4 15'34.22'
135.9 16' 173.63
' 141.8 1" 14.14
' 126.2 2" 22.75
' 128.4 3" 31.96
' 136.1 17" 26.17
' 199.7 18" 7o, 58
' 38.5 9' 24.3 1690.1610.1475.1400°1370.
1295.123011185.1135.995.9
80.91o1 840.730 33- Phosphorylcholine synthesis example 1 C42P(0)OCH2CH used in (B)
2Br 3. Og was dissolved in dry chloroform 10- and cooled on ice. To this were added a solution of 1.0 g (29 mmol) of the ether product obtained in 1'1 dissolved in 10 g of dry chloroform, and a solution of 1.0 g of dry triethylamine 4-1 dissolved in 10 g of dry chloroform, respectively. They were added dropwise simultaneously from separate funnels. After stirring the reaction mixture at room temperature for 20 hours, 10 - of a 5M aqueous potassium chloride solution was slowly added dropwise, and then 10 -
Stir for hours.
これにメタノール10mを加えた後、クロロホルム層を
集め、水で1回洗浄し、蒸発乾固した。After adding 10 m of methanol to this, the chloroform layer was collected, washed once with water, and evaporated to dryness.
残分を、さらに、五酸化リン上で減圧乾燥した後、乾燥
ブタノン15mに溶解した。この溶液をステンレス鋼製
耐圧反応管(20o−用)に仕込み、−25℃に冷却し
た後、乾燥トリメチルアミン15−を加えて密栓した。The residue was further dried under reduced pressure over phosphorus pentoxide and then dissolved in 15 ml of dry butanone. This solution was charged into a stainless steel pressure-resistant reaction tube (for 20°C), cooled to -25°C, and then dried trimethylamine 15- was added and the tube was tightly stoppered.
この反応混合物を60℃のポリエチレングリコール浴中
で9時間反応させた後生成した沈でんをろ集し、34−
乾燥ブタノンで洗浄した。This reaction mixture was reacted for 9 hours in a polyethylene glycol bath at 60°C, and the precipitate produced was collected by filtration and washed with 34-dry butanone.
こうして得た生成物をそのままメタノール70−に溶解
し、酢酸銀2.5グラムを加え、暗色において室温で1
.5時間攪拌した。ろ通抜、ろ液を蒸発に供し、残分を
減圧乾燥した後、シリカダルカラム(クロロホルム/メ
タノール/水−65/25/4 )で精製した。こうし
て所望のホスファチジルコリン型リン脂質化合物を得た
。The product thus obtained was dissolved directly in 70-methanol, 2.5 g of silver acetate was added, and the mixture was heated in the dark at room temperature for 1 hour.
.. Stirred for 5 hours. After filtering and draining, the filtrate was subjected to evaporation, and the residue was dried under reduced pressure, and then purified using a silica Dull column (chloroform/methanol/water - 65/25/4). In this way, the desired phosphatidylcholine type phospholipid compound was obtained.
収量0.60グラム(収率28チ)g
呈色あシ)
” C−NMRスペクトル(δ値):
炭素番号 δ(ppm) 炭素番号 δ(ppm
)1.3 64.3 7’ 19
9.82 77.0 8’ 3
8.54 59.0 9’ 2
4.46 54.4 14’ 2
4.91’ 116.5 15’
34.32’ 135.9 16’
173.73’ 141.8 1“
14.14’ 126.3 2“
22,75’ 128.4 3
“ 31.96’ 136.1 1
7“ 26.11735.1685.1615
.1470゜1410.1360.1240.1210
.1095.1080.975.925、45
窒素分析値:N1.80(1,79)
ただしく)内はC44H78N108P1の計算値。Yield: 0.60 grams (yield: 28 grams) C-NMR spectrum (δ value): Carbon number δ (ppm) Carbon number δ (ppm)
)1.3 64.3 7' 19
9.82 77.0 8' 3
8.54 59.0 9' 2
4.46 54.4 14' 2
4.91' 116.5 15'
34.32' 135.9 16'
173.73' 141.8 1"
14.14' 126.3 2"
22,75' 128.4 3
“ 31.96' 136.1 1
7" 26.11735.1685.1615
.. 1470°1410.1360.1240.1210
.. 1095.1080.975.925, 45 Nitrogen analysis value: N1.80 (1,79) The value inside is the calculated value for C44H78N108P1.
手続補正書
昭和58年11月16日
1、事件の表示
特願昭57−140963号
2、発明の名称
ホスファチジルコリン型リン脂質化合物3、補正をする
者
事件との関係 特許出願人
氏名 土 1) 英 俊
4、代理人
住所 東京都港区虎ノ門1丁目
26番5号第17森ビル
7、補正の内容
(1)明細書第18頁下から2ないし1行目にある「2
−ステアリル」を「2−〇−ステアリル」と訂正する。Procedural amendment dated November 16, 1981 1, Indication of the case, Patent Application No. 140963/1982, Name of the invention: Phosphatidylcholine type phospholipid compound 3, Person making the amendment Relationship to the case Patent applicant name Sat 1) English Shun 4, Agent address: 17 Mori Building 7, 1-26-5 Toranomon, Minato-ku, Tokyo Contents of amendment (1) “2” in the second or first line from the bottom of page 18 of the specification
-stearyl” is corrected to “2-〇-stearyl”.
(2)明細書第21頁5ないし6行目にある「2−ステ
アリル」を「2−O−ステアリル」と訂正する。(2) "2-stearyl" in lines 5 to 6 on page 21 of the specification is corrected to "2-O-stearyl."
(3)明細書第186行目にある「ホスホリルコリン」
を「ホスホコリン」と訂正する。(3) “Phosphorylcholine” on line 186 of the specification
is corrected to "phosphocholine".
(4)明細書第33頁下から1行目にある「2−ステア
リル」をr2−0−ステアリル」と訂正する。(4) "2-stearyl" in the first line from the bottom of page 33 of the specification is corrected to "r2-0-stearyl".
(5)明細書第18頁1行目にある「ホスホリルコリン
」を「ホスホコリン」と訂正する。(5) "Phosphorylcholine" on page 18, line 1 of the specification is corrected to "phosphocholine."
(6)明細書第36頁の表の右欄にある「炭素番号」の
下側にあるr17”Jの直下にr18”Jを挿入する。(6) Insert r18''J directly below r17''J under "Carbon Number" in the right column of the table on page 36 of the specification.
(7)明細書第36頁の表の右欄にある「σ(ppm)
」の下側にあるr26.IJの直下にr70.5Jを挿
入する。(7) "σ (ppm)" in the right column of the table on page 36 of the specification
” below r26. Insert r70.5J directly below IJ.
(8)明細書824頁下から16行目にある「参考例」
を「参考例1」と訂正する。(8) “Reference example” on page 824 of the specification, line 16 from the bottom
is corrected to "Reference Example 1".
(9)明細書第25頁4行目にある「であった。」と同
5行目にある「実施例2」との間に行を改めて以下の文
章を挿入する。(9) Insert a new line and insert the following sentence between "It was" on the 4th line of page 25 of the specification and "Example 2" on the 5th line.
「参考例 2
実施例1で得たリン脂質化合物20+agに0.05M
−リン酸緩衝水(pH7、0) 10mlを加え、窒素
雰囲気下で超音波処理 (50W 。"Reference Example 2 0.05M to the phospholipid compound 20+ag obtained in Example 1
- Add 10 ml of phosphate buffered water (pH 7, 0) and perform ultrasonic treatment (50 W) under a nitrogen atmosphere.
40℃で20分間)し、さらに40℃で1時間静置した
のち、30Wの低圧水銀ランプ(理工科学産業株式会社
製UVL−30LA)を用いて、窒素下に50℃で光源
から50鳳の距離から光照射した。重合の確認は、この
リポソーム水溶液151を」二記緩衝液10m1に稀釈
(リン脂質化合物濃度S
3.84xlOM)L、紫外吸収スペクトルを経時的に
測定することによっておこなった。反応に伴い、リン脂
質子ツマ−に基づく吸収(λ%αg26Lnm)が消失
し、新たに入−3X251 nmの吸収が生じた。A
wqa×の吸収は1時間後には完全に消失しており、重
合の完結を確認した。40°C for 20 minutes), and then left at 40°C for 1 hour. Using a 30W low-pressure mercury lamp (UVL-30LA manufactured by Riko Kagaku Sangyo Co., Ltd.), 50°C was heated under nitrogen at 50°C from the light source. Light was irradiated from a distance. Confirmation of polymerization was performed by diluting this liposome aqueous solution 151 in 10 ml of buffer solution (phospholipid compound concentration S: 3.84 x 1 OM) and measuring the ultraviolet absorption spectrum over time. As the reaction progressed, the absorption based on phospholipid molecules (λ%αg26Lnm) disappeared, and a new absorption at -3×251 nm was generated. A
Absorption of wqax completely disappeared after 1 hour, confirming completion of polymerization.
なお、生成重合体は、クロロホルム、メタノール等の有
機溶媒に不溶であった。Note that the produced polymer was insoluble in organic solvents such as chloroform and methanol.
参考例 3
すし
実施例1でたリン脂質化合物の重合をさらに確認するた
めに、エタノール中で酸化白金を触媒として室温で一時
間水素ガスを吹込んでビニル基の接触水素還元をおこな
った。得られた水素添加物と元のリン脂質化合物および
光重合物の各々の紫外吸収スペクトルを比較し、水素添
加物と光重合物の類似から、光重合物において重合によ
りビニル基が消失していることが判明した。次に、紫外
吸収スペクトルの測定結果を示す。Reference Example 3 Sushi In order to further confirm the polymerization of the phospholipid compound in Example 1, catalytic hydrogen reduction of vinyl groups was performed in ethanol using platinum oxide as a catalyst and blowing hydrogen gas at room temperature for one hour. Comparing the ultraviolet absorption spectra of the obtained hydrogenated product, the original phospholipid compound, and the photopolymerized product, it was found that the vinyl groups in the photopolymerized product disappeared due to polymerization due to the similarity between the hydrogenated product and the photopolymerized product. It has been found. Next, the measurement results of the ultraviolet absorption spectrum are shown.
9 εw+、x
)リン脂質 エタノ−27Bn+*(2xlO)、21
5n+w(0,9ヒ ル
5!10 204n層
1.48xlO水素添加 エタノール 251nm
(1,43z104) 。9 εw+,x
) Phospholipid ethano-27Bn+* (2xlO), 21
5n+w (0,9 hill
5!10 204n layer 1.48xlO hydrogenated ethanol 251nm
(1,43z104).
205nm 1.48xlO
光重合物 水 251nm(1,13xlO”
)202nm 1.23xlOII
参考例 4
紫外光照射による重合の難易度を実施例1のリン脂質化
合物と文献(III)に記載されたジエン型化合物との
間で比較した。各々100mgに水を加えて超音波処理
(50W、40℃で20分間)し、1%(wハ)濃度の
リポソーム水溶液を作り、参考例2と同一の条件で光照
射した。この水溶液の一部を採取し2.0xlOMとな
るように水で稀釈し、紫外吸収スペクトルを経時的に測
定した。文献(III)の化合物は入■0に261nm
の吸収が完全に消失するのに7時間を要するのに対し、
実施例1のリン脂質化合物にあっては、λwaX2.6
1nmの吸収の消失は2時間以内で完了した。J(10
)明細書第36頁最終行にある「計算値。」の次に行を
改めて以下の文章を挿入する。205nm 1.48xlO Photopolymerized water 251nm (1,13xlO”
) 202 nm 1.23xlOII Reference Example 4 The difficulty of polymerization by ultraviolet light irradiation was compared between the phospholipid compound of Example 1 and the diene type compound described in Document (III). Water was added to 100 mg of each and subjected to ultrasonication (50 W, 40°C for 20 minutes) to prepare a liposome aqueous solution with a concentration of 1% (w), and irradiated with light under the same conditions as in Reference Example 2. A portion of this aqueous solution was collected and diluted with water to 2.0xlOM, and the ultraviolet absorption spectrum was measured over time. The compound of literature (III) has a wavelength of 261 nm at 0
It takes 7 hours for the absorption of
In the phospholipid compound of Example 1, λwaX2.6
Disappearance of 1 nm absorption was completed within 2 hours. J(10
) Insert the following sentence in a new line after "Calculated value." on the last line of page 36 of the specification.
r参考例 5
実施例2で得たリン脂質化合物20mgを、参考例2と
同様にして重合させて重合化リポソームが得られること
を確認した。なお、生成重合体は、クロロホルム、メタ
ノール等の有機溶媒に不溶であった・
実施例 3
実施例1の(A−1)に従い、2−フェニル−5−m−
ジオキサノールと臭化ミリスチルとを反応させて合成し
た。収率29%。融点55.5−56℃。r Reference Example 5 It was confirmed that 20 mg of the phospholipid compound obtained in Example 2 was polymerized in the same manner as in Reference Example 2 to obtain a polymerized liposome. In addition, the produced polymer was insoluble in organic solvents such as chloroform and methanol. Example 3 According to (A-1) of Example 1, 2-phenyl-5-m-
It was synthesized by reacting dioxanol and myristyl bromide. Yield 29%. Melting point 55.5-56°C.
元素分析値(重量%):H(12,58)C(70,7
8)
(但し括弧内はC1ヮH,、O,の
計算値)
C−NMRスペクトルδ(ppm)値(’CD CI、
。Elemental analysis value (wt%): H(12,58)C(70,7
8) (However, the value in parentheses is the calculated value of C1ヮH,,O,) C-NMR spectrum δ (ppm) value ('CD CI,
.
TMS)
1.3 81.9 6 28.02
79.8 ?−15211,3−30,04
70,21622,7
531、!3 17 14.1質量スペクト
ル: M+1 =28!3(分子量288)(A−2)
2−ミリスチルオキシ−1,3−Pクロパンジオール1
.0gと実施例2の(B)で合成したp−(クロロカル
ボニルノナノイル)スチレンとを実施例2(C)の手法
に従って反応させて1−(p−ビニルベンソイルノナメ
イル)グリセロ−ミリスチルエーテル−2を合成した。TMS) 1.3 81.9 6 28.02
79.8? -15211,3-30,04
70,21622,7 531,! 3 17 14.1 Mass spectrum: M+1 = 28!3 (molecular weight 288) (A-2) 2-Myristyloxy-1,3-P clopanediol 1
.. 0g and p-(chlorocarbonylnonanoyl)styrene synthesized in Example 2 (B) according to the method of Example 2 (C) to produce 1-(p-vinylbensoylnonamyl)glycero-myristyl ether. -2 was synthesized.
収率36%。シリカゲルTLC:Rf=0.17 (ベ
ンゼン/エーテル=9/1)
(紫外吸収、ブロモチモールブルーによる呈色あり)
質量スペクトル:M=558(分子量558)石−NM
R(重クロロホルム、TMS):δ (ppm)
0.88(t、3H,−CH,); 1.26(S幅
広、 −(CH駄−);1、[l(t、2H,−COC
:H2−); 2.0B(t、 IH,−0H)2.
32(t、2H,−C)I2COO−)。Yield 36%. Silica gel TLC: Rf = 0.17 (benzene/ether = 9/1) (ultraviolet absorption, coloration by bromothymol blue) Mass spectrum: M = 558 (molecular weight 558) Stone-NM
R (deuterochloroform, TMS): δ (ppm) 0.88 (t, 3H, -CH,); 1.26 (S wide, -(CH-); 1, [l (t, 2H, -COC
:H2-); 2.0B(t, IH, -0H)2.
32(t,2H,-C)I2COO-).
2.82(t、2H,KO−COCH2−);3.5−
3.7(m、5H,−H(2−、−cH−) ;?
4、15(d、2H,−CH,QCニー) 。2.82 (t, 2H, KO-COCH2-); 3.5-
3.7 (m, 5H, -H(2-, -cH-);? 4, 15 (d, 2H, -CH, QC knee).
5.35,5.82,8.7(d、d、q、IH,IH
,IH,−CH=CH,) ;7、C9(d、d、2H
,2n、 −@−)”C−NMR(重クロロホルム、T
MS):δ(ppm)
3 82.0 14° 24.91’
118.5 15’ 34.22
’ 135.9 18’ 173.
73’ 141.8 1″ 14.
14’ 126.3 2″ 22.
75° 128.4 3” 31.
96’ 138.2 13” 2E
t、17’ 199.8 14″ 7
0.68’ 38.5
9’ 24.4
(B)raC−1−8−−ビニルベンゾイル)ノナノイ
ル−2−0−ミリスチル−グリセロ−3−ホスホコリン
実施例2の(D)の手法に従い、上記1−(p−ビニル
ベンソイルノナノイル)グリセロ−ミリスチルエーテル
−2のホスホコリン化をおこなうことによって合成した
。収率30%。5.35, 5.82, 8.7 (d, d, q, IH, IH
,IH,-CH=CH,);7,C9(d,d,2H
,2n, -@-)"C-NMR (deuterated chloroform, T
MS): δ (ppm) 3 82.0 14° 24.91'
118.5 15' 34.22
' 135.9 18' 173.
73' 141.8 1'' 14.
14' 126.3 2" 22.
75° 128.4 3” 31.
96' 138.2 13" 2E
t, 17' 199.8 14" 7
0.68' 38.5 9' 24.4 (B) raC-1-8--vinylbenzoyl)nonanoyl-2-0-myristyl-glycero-3-phosphocholine according to the procedure (D) of Example 2, the above It was synthesized by phosphocholination of 1-(p-vinylbensoylnonanoyl)glycero-myristyl ether-2. Yield 30%.
シリカゲルTLC:Rf=0.27 (クロロホルム/
メタノール/水= 85/25/4)(紫外吸収、ブロ
モチモールブルーによる呈色あり)
元素分析値(重量%): N tsgO(19,3)
(括弧内はC4o H2ONI O1? PIの計
算値)13C−NMR(重クロロホルム、TMS):δ
(ppm)
−2−品 δ m)5− −算 δ m4
59. 14°
24,91’ 118.3 15
’ 34.32’ 135゜8
16° 173.fi3 ”
141.8 1″
14.14’ 126.1 2″
22.85’ 128.3
3″ 31.96’
136.1 13”
28.07’ 199.8 1
4″ 70.58’ 38.5
5 86.49’
24.4 6 5
4.4IRスペクトル(クロロホルム溶液)CIIl:
2940、2870.1730.1B80.1810.
1470゜+410.1380.−1220.1180
.1090.975゜20
参考例 6
実施例3で得たリン脂質化合物20mgを、参考例2と
同様にして重合させて重合化リポソームが0
得られることを確認した。なお、生成重合体は、クロロ
ホルム、メタノール等の有機溶媒に不溶であった。」
1Silica gel TLC: Rf=0.27 (chloroform/
Methanol/water = 85/25/4) (ultraviolet absorption, coloring due to bromothymol blue) Elemental analysis value (wt%): N tsgO (19,3)
(In parentheses is C4o H2ONI O1? Calculated value of PI) 13C-NMR (deuterochloroform, TMS): δ
(ppm) -2-product δ m)5- -calculation δ m4
59. 14°
24,91' 118.3 15
'34.32' 135°8
16° 173. fi3”
141.8 1″
14.14' 126.1 2"
22.85' 128.3
3"31.96'
136.1 13”
28.07' 199.8 1
4″ 70.58’ 38.5
5 86.49'
24.4 6 5
4.4 IR spectrum (chloroform solution) CIII:
2940, 2870.1730.1B80.1810.
1470°+410.1380. -1220.1180
.. 1090.975°20 Reference Example 6 It was confirmed that 20 mg of the phospholipid compound obtained in Example 3 was polymerized in the same manner as in Reference Example 2 to obtain 0 polymerized liposomes. Note that the produced polymer was insoluble in organic solvents such as chloroform and methanol. ” 1
Claims (2)
示されるホスファチジルコリン型リン脂質化合物。(1) A phosphatidylcholine-type phospholipid compound represented by the general formula % (a positive integer of the formula %, and t is a positive integer of 13 to 21).
許請求の範囲第1項記載のリン脂質化金物。(2) The phospholipidated metal article according to claim 1, wherein t is 13#15, 17.19 or 21.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP14096382A JPS5931786A (en) | 1982-08-16 | 1982-08-16 | Phosphatidyl choline-type phospholipid compound |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP14096382A JPS5931786A (en) | 1982-08-16 | 1982-08-16 | Phosphatidyl choline-type phospholipid compound |
Publications (2)
Publication Number | Publication Date |
---|---|
JPS5931786A true JPS5931786A (en) | 1984-02-20 |
JPH0261957B2 JPH0261957B2 (en) | 1990-12-21 |
Family
ID=15280886
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP14096382A Granted JPS5931786A (en) | 1982-08-16 | 1982-08-16 | Phosphatidyl choline-type phospholipid compound |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPS5931786A (en) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS61129190A (en) * | 1984-11-27 | 1986-06-17 | Nippon Oil & Fats Co Ltd | Polymerizable glycerophospholipid |
US5091527A (en) * | 1988-03-04 | 1992-02-25 | Boehringer Mannheim Gmbh | Substrate for phospholipase |
US5466853A (en) * | 1992-07-23 | 1995-11-14 | Nof Corporation | Diester monomer, its polymer, water-containing soft contact lens, and processing solution for contact lens |
WO2003053984A1 (en) * | 2001-12-13 | 2003-07-03 | Gil-Ja Jhon | A process for preparing n-acylated lysophosphatidylcholine and a pharmaceutical composition for treatment of metabolic bone disease comprising said compounds |
-
1982
- 1982-08-16 JP JP14096382A patent/JPS5931786A/en active Granted
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS61129190A (en) * | 1984-11-27 | 1986-06-17 | Nippon Oil & Fats Co Ltd | Polymerizable glycerophospholipid |
JPH0455433B2 (en) * | 1984-11-27 | 1992-09-03 | Nippon Oils & Fats Co Ltd | |
US5091527A (en) * | 1988-03-04 | 1992-02-25 | Boehringer Mannheim Gmbh | Substrate for phospholipase |
US5466853A (en) * | 1992-07-23 | 1995-11-14 | Nof Corporation | Diester monomer, its polymer, water-containing soft contact lens, and processing solution for contact lens |
WO2003053984A1 (en) * | 2001-12-13 | 2003-07-03 | Gil-Ja Jhon | A process for preparing n-acylated lysophosphatidylcholine and a pharmaceutical composition for treatment of metabolic bone disease comprising said compounds |
CN1329403C (en) * | 2001-12-13 | 2007-08-01 | 钱吉子 | A process for preparing N-acylated lysophosphatidylcholine compounds and a pharmaceutical composition for treatment of metabolic bone disease comprising said compounds |
Also Published As
Publication number | Publication date |
---|---|
JPH0261957B2 (en) | 1990-12-21 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Akasaka et al. | Study on aromatic phosphines for novel fluorometry of hydroperoxides (I)-synthesis and spectral properties of diphenyl aryl phosphines and their oxides | |
Gaertner et al. | Removal of methyl groups from retinal controls the activity of bacteriorhodopsin | |
JPH0694444B2 (en) | Calcium specific fluorescent indicator dye | |
Huang et al. | Photocyclization of 2-(2'-hydroxyphenyl) benzyl alcohol and derivatives via o-quinonemethide type intermediates | |
JPS5931786A (en) | Phosphatidyl choline-type phospholipid compound | |
US20030134824A1 (en) | Beta-cyclodextrin dimers and phthalocyanines and uses thereof | |
Hansen et al. | Acridine-psoralen amines and their interaction with deoxyribonucleic acid | |
Moss et al. | Surface-specific cleavage of a cationic carbonate-functionalized vesicular surfactant | |
EP0251229B1 (en) | Polymerizable beta-glycerophospholipid and method for production thereof | |
Lala et al. | Orientation of the benzophenone group at various depths in bilayers | |
CN110642848B (en) | Synthesis and application of visible light and near-infrared fluorescent dye | |
JPS6191A (en) | Polymerizable phospholipid compound | |
JPS60116689A (en) | Phospholipid derivative | |
JPS61129192A (en) | Polymerizable phospholipid | |
Fatope et al. | Design, synthesis, and testing of potential antisickling agents. 10. 2, 2-Dimethylchroman-6-ylalkanoic acids | |
JP5354591B2 (en) | AMIDE COMPOUND HAVING PHOTOISOMERIZATION GROUP, ORGANIC NANOTUBE COMPRISING SELF-ASSEMBLING THE COMPOUND, AND METHOD FOR PRODUCING THE SAME | |
Elder | Microwave synthesis of tetraphenylcyclopentadienone and dimethyl tetraphenylphthalate | |
JPS6322541A (en) | Esters of sarsalate and guaiacol for treating inflammatory bronchial disease | |
Hori et al. | Stable 2-thianaphthalenes: synthesis and reactions with electrophiles | |
JPS6081192A (en) | Phospholipid compound | |
JPS58206540A (en) | Beta-cyclodextrin inclusion compound of ubidecarenone and its preparation | |
JPS6354385A (en) | Mixed acid-type polymerizable phospholipid derivative and production thereof | |
JPS60116690A (en) | Phospholipid derivative | |
JPS62108886A (en) | Phospholipid derivative | |
JPH0710874B2 (en) | Polymerizable phospholipid |